Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
SphK1 Double Nickase Plasmid (h): sc-401274-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SphK1 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- SphK1 Double Nickase Plasmid (h) and SphK1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SPHK1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SphK1 Antibody (G-11): sc-365401
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SphK1 Double Nickase Plasmid (h) | sc-401274-NIC | 20 µg | $410.00 | |||
SphK1 Double Nickase Plasmid (h2) | sc-401274-NIC-2 | 20 µg | $410.00 |
SPHK1 encodes sphingosine kinase 1 (SphK1), a lipid kinase that phosphorylates sphingosine to generate sphingosine-1-phosphate (S1P), a bioactive sphingolipid that influences cell survival, proliferation, migration, and inflammatory signaling. By shifting the balance between pro-apoptotic ceramide/sphingosine and pro-survival S1P, SphK1 participates in the sphingolipid rheostat and integrates inputs from growth factor and cytokine pathways. S1P produced by SphK1 can act intracellularly and through S1P receptors to modulate downstream programs including MAPK/ERK, PI3K/AKT, NF-κB, and angiogenic responses. Dysregulated SPHK1/S1P signaling has been linked to oncogenic phenotypes, fibrosis, and immune-mediated inflammation, making it a widely studied node in disease-relevant signaling networks.
SphK1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SPHK1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SPHK1. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt SPHK1 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of SPHK1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.